title: Identification of pharmacological modulators of HMGB1-induced inflammatory response by cell-based screening.
abstract: High mobility group box 1 (HMGB1), a highly conserved, ubiquitous protein, is released into the circulation during sterile inflammation (e.g. arthritis, trauma) and circulatory shock. It participates in the pathogenesis of delayed inflammatory responses and organ dysfunction. While several molecules have been identified that modulate the release of HMGB1, less attention has been paid to identify pharmacological inhibitors of the downstream inflammatory processes elicited by HMGB1 (C23-C45 disulfide C106 thiol form). In the current study, a cell-based medium-throughput screening of a 5000+ compound focused library of clinical drugs and drug-like compounds was performed in murine RAW264.7 macrophages, in order to identify modulators of HMGB1-induced tumor-necrosis factor alpha (TNFα) production. Clinically used drugs that suppressed HMGB1-induced TNFα production included glucocorticoids, beta agonists, and the anti-HIV compound indinavir. A re-screen of the NIH clinical compound library identified beta-agonists and various intracellular cAMP enhancers as compounds that potentiate the inhibitory effect of glucocorticoids on HMGB1-induced TNFα production. The molecular pathways involved in this synergistic anti-inflammatory effect are related, at least in part, to inhibition of TNFα mRNA synthesis via a synergistic suppression of ERK/IκB activation. Inhibition of TNFα production by prednisolone+salbutamol pretreatment was also confirmed in vivo in mice subjected to HMGB1 injection; this effect was more pronounced than the effect of either of the agents administered separately. The current study unveils several drug-like modulators of HMGB1-mediated inflammatory responses and offers pharmacological directions for the therapeutic suppression of inflammatory responses in HMGB1-dependent diseases.
authors.affiliation: Department of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, United States of America.
authors.forename: Domokos
authors.initials: D
authors.lastname: Gerö
authors.forename: Petra
authors.initials: P
authors.lastname: Szoleczky
authors.forename: Katalin
authors.initials: K
authors.lastname: Módis
authors.forename: John P
authors.initials: JP
authors.lastname: Pribis
authors.forename: Yousef
authors.initials: Y
authors.lastname: Al-Abed
authors.forename: Huan
authors.initials: H
authors.lastname: Yang
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Chevan
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Billiar
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Tracey
authors.forename: Csaba
authors.initials: C
authors.lastname: Szabo
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000420:Albuterol / Q000494:pharmacology; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002395:Catecholamines / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology; D015536:Down-Regulation; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D004734:Energy Metabolism; D015870:Gene Expression / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology; D024243:HMGB1 Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D010766:Phosphorylation; D011239:Prednisolone / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000318:Adrenergic beta-Agonists; D000893:Anti-Inflammatory Agents; D002395:Catecholamines; D005938:Glucocorticoids; D024243:HMGB1 Protein; C582808:HMGB1 protein, mouse; D018836:Inflammation Mediators; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D011239:Prednisolone; D000420:Albuterol